Chinese traditional medicine composition for treatment of avian influenza, method for preparation and application thereof

ABSTRACT

A Chinese traditional medicine composition for prevention and treatment of avian influenza is provided. Its active ingredients are consisted of: herba houttuyniae 21.4 wt. %, Caulis lonicerae japonicae 17.9 wt. %, radix isatidis 14.3 wt. %, male fern rhizome 10.7 wt. %, radix et rhizoma sophorae tonkinensis 7.1 wt. %, radix angelicae dahuricae 7.1 wt. %, rhizoma paridis 7.1 wt. %, herba artemisiae annuae 7.2 wt. %, and rhizoma iridis tectoris 7.2 wt. %. This Chinese traditional medicine composition could be made into any regular medicinal types, for example, oral solutions, tablets, capsules, pellets and injection solution, etc. Modified preparation methods for this Chinese traditional medicine composition, as well as the use of the same in preparation of medicines and health foods for prevention or treatment of avian influenza were also provided in present invention. This Chinese traditional medicine composition could effectively prevent and treat avian influenza. It was a safer, more easily-obtained, and inexpensive anti-avian influenza drug.

FIELD OF THE INVENTION

The present invention relates to a Chinese traditional medicinecomposition for prevention and treatment of Avian Influenza (AI), amethod for preparation, and application thereof. More specifically, thepresent invention relates to a Chinese traditional medicine compositionfor prevention and treatment of AI that is made from herb, a method forpreparing the same, and application thereof.

BACKGROUND OF THE INVENTION

AI is an avian acute contagious symptom caused by type A influenzavirus. The affected animals include geese, chickens, ducks, pigeons andsome wild birds. The research on contagious diseases has indicated thatAIV (Avian Influenza Virus) is highly pathogenic and contagious. AI wasdefined by World Organization for Animal Health as a type A infection,also known as fowl plague or European plague. There are three types ofAI in relation to the pathogen. These include highly pathogenic, lowpathogenic and non-pathogenic. No obvious symptoms were caused bynon-pathogenic AI. However, some virus antibody was generated in theinfected birds. The symptoms caused by low pathogenic AI includedmarginal respiratory symptom, reduced food intake, reduced eggproduction, and a few deaths. The highly pathogenic AI could cause themost serious symptoms with a very high death rate. Humans have not yethad effective methods for prevention and treatment of high pathogenicAI, which was found in 1878. Sterilization, separation and flockslaughtering were applied to prevent further spreading. The infectedflock had an extremely high death rate which brought serious financialloss.

Naturally, the correlation between AI and human flu happened throughintermediate hosts such as pigs, horses, dolphins, and other mammals.However, the antigenic characteristics of the highly pathogenic AIVchanged frequently, which was also called antigenic shift and antigenicdrift. In addition, its intermediate hosts increased continuously. Thehighly pathogenic AIV nowadays could infect humans directly. The firstcase of inter-species infection was in Hong Kong, China, in 1997, andagain on China's mainland and Hong Kong in 1999. It was induced by H₉N₂AIV and caused some mortality. It broke the rule in which human andother mammals could only be infected by H₁, H₂, and H₃ influenza virus.H₅ and H₉ are the newest members of the influenza virus family, whichbring new topic for public health. Avian Influenza has broken out inmany countries and it has become a major threat to the human health andthe safety of global economy.

H₁, H₂, and H₃ were the only influenza virus's found in humans for along time. All the research, vaccines, and anti-viral medications wereaimed towards these three viruses. Human beings had neither immunitiesnor effective medications against the new outbreak of H₅, H₁, and H₇.Serious symptoms, including fever, muscle soreness, and chills, could becaused in humans by highly pathogenic AIV infections. Its complicationscould even cause death.

The current major medications for anti-influenza virus includeneuralminidase inhibitors (such as Zanamivir, Oseltamivir, etc.), ionchannel blockers (such as Amantadine and rimantadine), and nucleotidemedications (such as tribavirim, that is, ribavirin or virazole). Ionchannel blockers were considered as the best medications for preventionand treatment of influenza viruses.

However, the above mentioned medications have shown limitations:

(1) The neuralminidase inhibitors could effectively restrain type A andB influenza viruses. However, it is relatively expensive, which limitsits extension. For example, Tamiflu made in Roche Pharmaceutical Corp.was also called Oseltamivirphosphat, i.e., phosphate of Oseltamivir((3R,4R,5S)-4-acetylamino-5-amino-3(1-ethylpropoxy)-1-cyclohexene-1-carboxylicacid ethyl ester, with CASRN of 196618-13-0) with CASRN of 204255-11-8.This medicine should be taken twice daily with a dosage of 75 mg eachtime. Thus 60 RMB was needed per day for medicine. 300 RMB was requiredfor one 5-day treatment phase. Furthermore, Roche Pharmaceutical Corp.refused to release the patent for Tamiflu even after 2017, citingjustifications such as the complexity of production and maintaining highquality standards. Thus the production of Tamiflu was in short supply.

However, significant side effects have been found with Tamiflu,including hallucinations, abnormal behaviors and other psychological andneural symptoms. Drug resistance has also been observed in some of thepatients.

(2) Ion channel blockers could effectively resist type A influenzavirus. However, it is poisonous to the nervous system and could inducedrug resistance if taken for a long period of time. It also has noeffect on type B influenza virus.

(3) Tribavirin (virazole) could effectively treat the infection causedby RNA and DNA viruses. However, this compound has teratogenic effectwhich limits its application in clinical practices.

In order to find effective and safe treatment for influenza viruses,many scientists turned to Chinese traditional herbs for help. Forexample, a Chinese traditional medicine composition which could preventchicken, duck avian influenza, was introduced in articleCN200510011432.3 from Huanan Agricultural University. However, thisChinese traditional medicine composition was not very effective in avianinfluenza treatment, and thus it cannot meet the requirement forinhibiting the outbreak of AI.

Therefore, a safe, effective, inexpensive medicine which could not onlyprevent but also treat avian influenza is in great need.

SUMMARY OF THE INVENTION

One purpose of the present invention is to provide a Chinese traditionalmedicine composition for prevention and treatment of avian influenza.

Another purpose of the present invention is to provide a preparationmethod for the Chinese traditional medicine composition of the presentinvention.

Yet the other purpose of this invention is to provide uses of theChinese traditional medicine composition of the present invention inpreparation of medicines which are used in prevention and treatment ofavian influenza.

A further purpose of this invention is to provide uses of the Chinesetraditional medicine composition of the present invention in preparationof health goods which are used in prevention and treatment of avianinfluenza.

In one embodiment of the present invention, the Chinese traditionalmedicine composition which could effectively prevent and treat avianinfluenza comprises active ingredients and/or medical acceptable vector,wherein the active ingredients substantially includes: herba houttuyniae4.7 wt. %-70.0 wt. %, Caulis lonicerae japonicae 4.7 wt. %-46.7 wt. %,radix isatidis 2.3 wt. %-42.0 wt. %, radix et rhizoma sophoraetonkinensis 1.2 wt. %-35.0 wt. % and other herbs for removing fever anddetoxication.

The above mentioned herbs are Male Rern Rhizome, Radix angelicaedahuricae, Herba Andrographic, Dandelion, Natural Indigo,Sargentgloryvine Stem, Chinese Licorice, or the like.

The active ingredients of the above mentioned Chinese traditionalmedicine composition could also include herba houttuyniae 4.7 wt. %-70.0wt. %, Caulis lonicerae japonicae 4.7 wt. %-46.7 wt. %, radix isatidis2.3 wt. %-42.0 wt. %, and radix et rhizoma sophorae tonkinensis 1.2 wt.%-35.0 wt. %.

In another embodiment of the present invention, the Chinese traditionalmedicine composition which could effectively prevent and treat avianinfluenza comprises active ingredients and/or medical acceptable vector,wherein the active ingredients substantially includes herba houttuyniae4.7 wt. %-58.4 wt. %, Caulis lonicerae japonicae 4.7 wt. %-46.7 wt. %,radix isatidis 2.3 wt. %-42.0 wt. %, radix et rhizoma sophoraetonkinensis 1.2 wt. %-35.0 wt. %, male fern rhizome 1.9 wt. %-37.3 wt.%, radix angelicae dahuricae 1.2 wt. %-35.0 wt. %, and other herbs forfever reduction and detoxification.

The above mentioned herbs are rhizoma paridis, herba artemisiae annuae,Java brucea fruit, herba andrographis, dandelion, mint, platycodongrandiflorus, Chinese licorice, or the like.

The active ingredients of the above mentioned Chinese traditionalmedicine composition could also include herba houttuyniae 4.7 wt. %-58.4wt. %, Caulis lonicerae japonicae 4.7 wt. %-46.7 wt. %, radix isatidis2.3 wt. %-42.0 wt. %, radix et rhizoma sophorae tonkinensis 1.2 wt.%-35.0 wt. %, male fern rhizome 1.9 wt. %-37.3 wt. %, and radixangelicae dahuricae 1.2 wt. %-35.0 wt. %.

In another embodiment of the present invention, the Chinese traditionalmedicine composition which could effectively prevent and treat avianinfluenza comprises active ingredients and/or medical acceptable vector,wherein the active ingredients is consisted of herba houttuyniae 9.3 wt.%-46.7 wt. %, Caulis lonicerae japonicae 9.3 wt. %-37.3 wt. %, radixisatidis 4.7 wt. %-35.0 wt. %, male fern rhizome 2.3 wt. %-28.0 wt. %,radix et rhizoma sophorae tonkinensis 2.3 wt. %-23.3 wt. %, radixangelicae dahuricae 2.3 wt. %-23.3 wt. %, rhizoma paridis 2.3 wt. %-23.3wt. %, herba artemisiae annuae 2.3 wt. %-23.3 wt. %, and rhizoma iridistectoris 2.3 wt. %-23.3 wt. %.

The preferred components of the active ingredients of the abovementioned Chinese traditional medicine composition are herba houttuyniae14.0 wt. %-35.0 wt. %, Caulis lonicerae japonicae 11.7 wt. %-23.3 wt. %,radix isatidis 9.3 wt. %-21.0 wt. %, male fern rhizome 7.0 wt. %-18.7wt. %, radix et rhizoma sophorae tonkinensis 4.7 wt. %-11.7 wt. %, radixangelicae dahuricae 4.7 wt. %-11.7 wt. %, rhizoma paridis 4.7 wt. %-11.7wt. %, herba artemisiae annuae 4.7 wt. %-11.7 wt. %, and rhizoma iridistectoris 4.7 wt. %-11.7 wt. %.

The more preferred components of the active ingredients of the abovementioned Chinese traditional medicine composition are herba houttuyniae21.4 wt. %, Caulis lonicerae japonicae 17.9 wt. %, radix isatidis 14.3wt. %, male fern rhizome 10.7 wt. %, radix et rhizoma sophoraetonkinensis 7.1 wt. %, radix angelicae dahuricae 7.1 wt. %, rhizomaparidis 7.1 wt. %, herba artemisiae annuae 7.2 wt. %, rhizoma iridistectoris 7.2 wt. %.

The Chinese traditional medicine composition of the present inventioncould be readily made into any form of drug by conventional methods. Forexample, the raw materials of the present invention could be ground intopowder, mixed and taken with water. The raw materials of the presentinvention could also be boiled and concentrated into medicines. In orderto maximize the effect of all the raw materials, the herba houttuyniaewas preferably extracted, for example, by using ethanol. However, thesecould not be used to limit the scope of the present invention.

The preferred Chinese traditional medicine composition comprises herbahouttuyniae, caulis lonicerae japonicae, radix isatidis and male fernrhizome, etc., could be prepared in the following steps:

The above mentioned four herbs were obtained in prescription amounts.The herba houttuyniae was boiled in water for 10˜40 minutes. Thesolution was filtered and concentrated into a clear cream with a densityof 1.05-1.20. The cream was then cooled and ethanol was added until theamount of ethanol was 50˜80%. It was then allowed to sit for 24-48 hoursfollowed by filtration. The collected ethanol was concentrated and aclear cream of density 1.05-1.25 was obtained. The other three herbs,including caulis lonicerae japonicae, radix isatidis and male fernrhizome, were boiled for 2 hours in water twice. The herbs were filteredand the filtrate was concentrated until a density of 1.40 was reached.Following filtration the obtained filtrate was re-concentrated into aclear cream with a density of 1.45. The herba houttuyniae clear creamwas then added and mixed. The mixture was continuously concentrated tothick cream with a density of 1.10 or greater. Finally, the thick creamand powdered sucrose were mixed in a 1:3˜8 ratio. The mixture was thenmade into pellets and packed.

The raw materials in the above mentioned preparation process could alsobe prepared with male fern rhizome and radix angelicae dahuricae, orwith herba andrographic, dandelion, natural indigo, sargentgloryvinestem or Chinese licorice. The detailed preparation steps are as follows:

Herba houttuyniae, caulis lonicerae japonicae, radix isatidis, radix etrhizoma sophorae tonkinensis, male fern rhizome and radix angelicaedahuricae were obtained in prescription amount. The herba houttuyniaewas boiled in water for 10˜40 minutes, filtered, and concentrated into aclear cream with a density of 1.20. The cream was cooled and ethanol wasadded until the amount of ethanol was 50˜80%. It was allowed to sit for24-48 hours followed by filtration. The ethanol was collected from thefiltrate and concentrated to obtain a clear cream with a density of1.25. The other 5 herbs were boiled in water twice for 2 hours eachtime. The mixture was then filtered and concentrated to a density of1.08. Filtered again, the filtrate was concentrated into a clear creamwith a density of 1.10, and mixed with the obtained herba houttuyniaeclear cream. The mixture was continuously concentrated to obtain a thickcream with a density of 1.26 or above. Finally, the thick cream andpowdered sucrose were mixed in a 1:3˜8 ratio. The mixture was then madeinto pellets and packed.

The Chinese traditional medicine composition of the present inventionconsisting of herba houttuyniae, caulis lonicerae japonicae, radixisatidis, radix et rhizoma sophorae tonkinensis, male fern rhizome,radix angelicae dahuricae, rhizoma paridis, herba artemisiae annuae, andrhizoma iridis tectori, could be prepared in the following steps:

All 9 herbs mentioned above were obtained in prescription amount. Theherba houttuyniae was boiled in water for 10˜40 minutes, filtered, andconcentrated to a clear cream with a density of 1.05. The cream wascooled and ethanol added until the amount of ethanol was 50˜80%. It wasallowed to sit for 24 hours then filtered. The ethanol was thencollected from the filtrate and concentrated to obtain a clear creamwith a density of 1.25. The other 8 herbs were boiled in water twice for2 hours each time. The mixture was then filtered and concentrated to adensity of 1.20. Filtered again and the filtrate was concentrated toobtain a clear cream with a density of 1.10-1.25, then mixed with theherba houttuyniae clear cream. The mixture was concentrated to obtain athick cream with a density of 1.26 or above. Finally, the thick creamand powder sucrose were mixed in a proportion of 1:3˜8. The mixture wasthen made into pellets and packed.

One or more medical acceptable vectors could be added to the Chinesetraditional medicine composition of the present invention. For example,regular accessory agents include diluting agents, excipients, fillers,adhesive agents, moisturizing agents, disintegrant agents, absorptionenhancers, surfactants, absorbing vectors, lubricants, and the like.

The Chinese traditional medicine composition of the present inventioncould be taken orally, applied through venous injection or mucousmembranes. It could be made into tablets, capsules, powder, pellets,pastille, suppositories, oral solutions, pasteurized gastroentericsuspension injections, small or large amounts of injection, frozen powerinjections, pasteurized powder injections or the like. All of theabove-mentioned medicines could be prepared using regular methods in theart.

In order to treat the infected animals or enhance the animal immunitiesagainst avian influenza, the Chinese traditional medicine composition ofthe present invention could be added to the animal forage as anadditive.

The effective application amount of the Chinese traditional medicinecomposition or additive of the present invention was 20-80 mg/kg bodyweight per day. The preferred application amount was 30-65 mg/kg bodyweight per day.

The raw materials of the Chinese traditional medicine composition of thepresent invention were natural plants. The Chinese traditional medicinecomposition could significantly resist the avian influenza viruses. Theprepared medicines or additives were safe, effective, and provided newmethods for prevention and treatment of avian influenza.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows an optical microscopic photograph of the cells of controlgroup in Experiment Example 3, while the culture of cells was stopped.FIG. 1A shows a positive control group without medicine. FIG. 1B shows acontrol group applied with 0.284 ug/ml of Tamiflu, and a large amount ofcells are infected. FIG. 1C shows a control group applied with 2.84ug/ml of Tamiflu, and most of cells are infected. FIG. 1D shows acontrol group applied with 28.4 ug/ml of Tamiflu, wherein viruses waskilled, but there are many unrelated cells (killed).

FIG. 2 shows optical microscopic photograph of the cells of controlgroup in Experiment Example 3, while the culture of cells was stopped.FIG. 2A shows a positive control group without medicine. FIG. 2B showsthe photograph when applied with 12.5 ug/ml of the Chinese traditionalmedicine composition of the present invention, and a large amount ofcells are infected. FIG. 2C shows photograph when applied with 25 ug/mlof the Chinese traditional medicine composition of the presentinvention, and a large amount of cells are infected. FIG. 2D shows thephotograph when applied with 50 ug/ml of the Chinese traditionalmedicine composition of the present invention, wherein the viruses arekilled, and only a few cells are infected. FIG. 2E shows the photographwhen applied with 100 ug/ml of the Chinese traditional medicinecomposition of the present invention, wherein all viruses are killed,and there are a few unrelated cells (killed). FIG. 2F shows thephotograph when applied with 200 ug/ml of the Chinese traditionalmedicine composition of the present invention, wherein all viruses arekilled, but some cells are killed.

DETAIL DESCRIPTION OF EMBODIMENTS OF THE INVENTION

The following Experiment Examples is to further describe the benefit ofthe Chinese traditional medicine composition of the present inventionover the current existing anti-avian influenza medicines. The followingExperiment Example includes the cytological and zoological experimentsfor the Chinese traditional medicine composition of the presentinvention.

Experiment Example 1

The protective effect of the Chinese traditional medicine composition ofthe present invention for the flocks infected with avian influenzaviruses.

The chickens used in this experiment were healthy 50-day old yellowchicken, which were purchased from the experimental chicken farm ofZhongshan University. The Chinese traditional medicine composition ofthe present invention (which is the pellet in Example 5), was providedby the Lizhu Pharmaceutical Group, Inc. The Rimantadine, one of thecontrol medicines, was purchased from Zhejiang Yukang PharmaceuticalCorp. The other control medicine was prepared according to China Patent200510011432.3. The virus was H5 serum type avian influenza virus(1.0×10⁻⁶).

Experimental Process:

190 healthy yellow chickens of 50-days old were randomly grouped into 5groups. 30 were in the normal control group, the other groups have 40yellow chickens each. Each chicken was injected in the abdomen with 0.2ml of avian influenza virus diluted in chicken embryo amniotic fluidwith a concentration of 0.5×10⁻⁵. Each group was fed with medicine thesame day infected with AIV, except the healthy control group andinfected control group, which were fed with physiological saline. Themedicine or physiological saline was applied once per day continuouslyfor 8 days. Group A was the healthy control group. Group B was appliedwith 10 mg/day the Chinese traditional medicine composition of thepresent invention. Group C was applied with 10 mg/day Rimantadine, oneof the control medicines. Group D was applied with 10 mg/day of themedicine prepared according to China Patent 200510011432.3. Group E wasthe infected control group. The results are shown in the following table1:

TABLE 1 Number of Rate of Effective Group Number Treatment method deathdeath rate A 30 Not infected 0 — — B 40 Infected, medicine 1  2.5% 97.5%applied C 40 Infected, medicine 23 57.5% 42.5% applied D 40 Infected,medicine 16   40%   60% applied E 40 Infected, medicine 37 92.5% — notapplied

The above table shows that the infected chickens were much moreeffectively protected by the Chinese traditional medicine composition ofthe present invention than medicine of China Patent 200510011432.3 andRimantadine.

Experiment Example 2

This was an experiment for treatment of viral pneumonia caused by H₇avian influenza virus.

The lab animals were the Kunming mice provided by the experimentalanimal lab of Haerbing Medical University. The mice were injected withH₅N₁ viruses. The Chinese traditional medicine composition injections inthis experiment were provided by the Lizhu Pharmaceutical Group, Inc.

Experimental Process:

Each of the experimental groups and control groups had 17 mice. Controlgroup 1 was injected in the abdomen with 15 ml/kg of physiologicalsaline. Control group 2 was injected in the abdomen with 15 ml/kg ofZedoray Tumeric Oil injection. Control group 3 was injected in theabdomen with 15 ml/kg of ShuangSuLian injection which was often used inthe treatment of pneumonia. Experimental group 1 was injected in theabdomen with 15 ml/kg of injection solution which was made fromhouttuyniae, caulis lonicerae japonicae, radix isatidis and male fernrhizome. Experimental group 2 was injected in the abdomen with 15 ml/kgof injection solution which was made from houttuyniae, caulis loniceracjaponicae, radix isatidis, male fern rhizome, and Chinese licorice.Experimental group 3 was injected in the abdomen with 15 ml/kg ofinjection solution which was made from houttuyniae, caulis loniceraejaponicae, radix isatidis, male fern rhizome, tokin sophora root, andradix angelicae dahuricae. All medicines were applied continuously for10 days. Mice in all groups were killed after 11 days and samples werecollected for testing. The results are shown in Table 2:

TABLE 2 Significant Group effective effective Not effective DeathExperimental group 1 9 4 4 3 Experimental group 2 8 3 6 5 Experimentalgroup 3 12 4 1 2 Control group 1 0 2 15 13 Control group 2 3 5 9 8Control group 3 2 3 12 10 Note: “Significantly effective” means thesymptoms were greatly relieved, including normal body temperature and agreatly reduced lung vesicular breath sound. “Effective” means thesymptoms were relieved, including normal body temperature and slightlyreduced lung vesicular breath sound. “Not effective” means the symptomswere not relieved at all or more severe.

Table 2 shows that the Chinese traditional medicine composition used inexperimental groups 1, 2, and 3 had a much more positive effect comparedto control groups 1, 2, and 3. The Chinese traditional medicinecomposition significantly improved lung circulation, reduced aggregationof blood platelets, reduced lung infection, healed lung damage, andresisted H₇ avian influenza virus. The Chinese traditional medicinecomposition in Experimental group 3 showed a much more positive effectthan in control groups 1 and 2.

Experiment Example 3

This experiment was the in vitro cytological research on the resistanceof the Chinese traditional medicine composition of the present inventionagainst highly pathogenic AI.

The medicines used in this experiment were the pellets made in Example7. The control medicine was Tamiflu, that is, Oseltamivir phosphate madeby Roche Corp. The cells used in this experiment were MDCK cells (dogkidney cells), which were provided by the Microbiology Department ofMedicine Faculty of the University of Hong Kong. The virus used in thisexperiment was H₅N₁ (geno type E), which was also provided by theMicrobiology Department of Medicine Faculty of the University of HongKong.

Experimental Process:

-   -   (1) Cell toxicity test: the medicine concentration (CC₅₀) which        induced pathological changes in 50% of the cells was tested        according Mosmann rules (1983).    -   (2) Anti-virus test: a certain amount of viral suspension was        put into bottles containing cell cultures. 8 bottles were placed        parallel for 45 minutes to let the viruses attach to the cells.        Cultural solutions lacking or with differing concentrations of        medicines were put into the bottles. The solutions were        incubated at 37° C. for 20 hours. The cultures were terminated        when 50% of the cells showed pathological changes in the bottle        with respect to control cells. Photographs were taken. The        results are shown in Table 3:

TABLE 3 Concentration of the Virus medicine (ug/ml) MedicineExperimental group H5N1(genotype E) 12.5 +8/8 Experimental groupH5N1(genotype E) 25 +8/8 Experimental group H5N1(genotype E) 50 +2/8Experimental group H5N1(genotype E) 100 +0/8 Experimental groupH5N1(genotype E) 200 +0/8 Positive control group Tamiflu H5N1(genotypeE) 0.284 +8/8 Tamiflu H5N1(genotype E) 2.84 +2/8 Tamiflu H5N1(genotypeE) 28.4 +0/8 Note: + means non-resistance, negative control cells werenormal cells.

Table 3 shows that the Chinese traditional medicine compositioninjection solution could effectively resist the avian influenza viruswhen the concentration of the active ingredients was 50 ug/ml or above.

The preparation methods of the Chinese traditional medicine compositionof the present invention are described as follows:

Example 1 Preparation of Large Volume Injections of the ChineseTraditional Medicine Composition of the Present Invention

First, the following herbs were obtained: 2000 g of herba houttuyniae,400 g of caulis lonicerae japonicae, 1500 g of radix isatidis, 284 g ofmale fern rhizome, 100 g of radix et rhizoma sophorae tonkinensis, 100 gof radix angelicae dahuricae, 100 g rhizoma paridis, 100 g of herbaartemisiae annuae, and 100 g of rhizoma iridis tectori. The herbs wereadded with 8× water, soaked for 3 hours, then boiled and extracted twicefor 1 hour each time. The extracts were combined, filtered, andconcentrated under vacuum to a density of 1.25 at 65° C. They were thencooled and mixed with ethanol until 70%, then allowed to sit. Thesupernatant was collected, and the ethanol was removed from the liquid.The obtained solution was then concentrated to a density of 1.15 at 65°C. and counter-current extracted using 2× ethyl acetate. The extractswere combined and the ethyl acetate was removed. 50% of the injectionwater was added. The liquid was then boiled, cooled, and filtered,followed by mixing with suitable amount of injection water. The solutionwas then mixed with 0.08% of emulsifier (T-80) followed by addition of40% of NaOH until a pH of 8.0 was reached. NaCl was added to thesolution followed by injection water to obtain the isotonic solution. Atlast, the liquid was filtered through 0.22 μm microvoid filter film,sealed, pasteurized, lamp-inspected, and packed.

Example 2 Preparation of Small Volume Injections of the ChineseTraditional Medicine Composition of the Present Invention

First, the following herbs were obtained: 400 g of herba houttuyniae,1600 g of caulis lonicerae japonicae, 200 g of radix isatidis, 100 g ofmale fern rhizome, 1000 g of radix et rhizoma sophorae tonkinensis, 200g of radix angelicae dahuricae, 200 g rhizoma paridis, 384 g of herbaartemisiae annuae, and 200 g of rhizoma iridis tectori. All herbs wereboiled, filtered, concentrated in a vacuum chamber, added to ethanol,allowed to sit, filtered and concentrated under vacuum until 9 gmedicine/ml were obtained. The solution was diluted in water, stored ina fridge, filtered, concentrated under vacuum, and separated in amacroporous resin which was washed with water, NaOH solution, 35 wt % ofethanol, and 75 wt % of ethanol. The eluate was collected, diluted withwater, combined with L-Arginase, adjusted until a pH 7.5 was reached,followed by addition of active carbon. The eluate was then heated,filtered, diluted in the same volume of water, followed by addition ofL-Arginase until a concentration of 4 wt % was reached.

Example 3 Preparation of Frozen-Dry Powder Injections of the ChineseTraditional Medicine Composition of the Present Invention

First, the following herbs were obtained: 584 g of herba houttuyniae,500 g of caulis lonicerae japonicae, 400 g of radix isatidis, 300 g ofmale fern rhizome, 500 g of radix et rhizoma sophorae tonkinensis, 500 gof radix angelicae dahuricae, 500 g rhizoma paridis, 500 g of herbaartemisiae annuae, and 500 g of rhizoma iridis tectori. All herbs wereadded to 3200 ml of injection water, soaked for 1 hour, added to 6400 mlof injection water, boiled twice for 1.5 hours each time, and filtered.The filtrate was combined and the extract composition was obtained. 10 gof the extract composition was diluted in injection water, and thenadded to 2.25 g of NaCl followed by injection water until a volume of250 ml was reached. Injection carbon was added and the solution wasboiled for 20 minutes, cooled, filtered, and pasteurized. The finalfrozen powder injection was then packed in low-boro glass ampoules,cooled, and dried.

Example 4 Preparation of Small Volume Injections of the ChineseTraditional Medicine Composition of the Present Invention

First, the following herbs were obtained: 200 g of herba houttuyniae,200 g of caulis lonicerae japonicae, 100 g of radix isatidis, 1279 g ofmale fern rhizome, 1500 g of radix et rhizoma sophorae tonkinensis, and1500 g of radix angelicae dahuricae. All herbs were added to water,boiled, and filtered. The mixture was concentrated under vacuum, addedto ethanol, allowed to sit, then filtered again. The mixture was againconcentrated under vacuum until 11 g medicine/ml was obtained. Themedicine was diluted in water, stored in a fridge, filtered a thirdtime, concentrated under vacuum, and separated in macroporous resinwhich was washed with water, NaOH solution, 55 wt % of ethanol, and 65wt % of ethanol. The eluate was collected, diluted, added L-Arginase,adjusted until a pH 7.0 was reached, added active carbon, heated, andfiltered. The final product was obtained by diluting of the eluate withthe same volume of water.

Example 5 Preparation of Pellets of the Chinese Traditional MedicineComposition of the Present Invention

First, the following herbs were obtained: 2500 g of herba houttuyniae,1000 g of caulis lonicerae japonicae, 400 g of radix isatidis, 784 g ofmale fern rhizome, 50 g of radix et rhizoma sophorae tonkinensis, and 50g of radix angelicae dahuricae. All herbs were added to 70 wt. % ofethanol, soaked overnight, refluxed to extract twice for 1.5 hours eachtime, and filtered. The residue I and solution I were collected. Theresidue I was put into a vacuum concentrating pot, and recovered undervacuum until no more ethanol remained, and concentrated into a thickcream.

Starch was dried, sieved. Mannitol was sieved, and allowed to sit. 500 gof the thick cream was mixed with 500 g mannitol and 150 g starch. Themixture was then made into pellets on the gramulator with steel mesh,dried on an ebullated bed, modified with 8 mesh screener, tested forcontent and water, packed with 7 g per bag, sealed and stored in a cooldry place.

Example 6 Preparation of Tablets of the Chinese Traditional MedicineComposition of the Present Invention

First, the following herbs were obtained: 3000 g of herba houttuyniae,1134 g of caulis lonicerae japonicae, 100 g of radix isatidis, and 50 gof radix et rhizoma sophorae tonkinensis. All herbs were added with 3200ml injection water, soaked for 1 hour, added with 3200 ml injectionwater again, boiled twice with 1 hour each time, and filtered. Theresidues were discarded. The solution was put in triple effectevaporator and concentrated until a density of 1.08 (70° C.) wasreached, and cooled to room temperature. 95% ethanol was added slowlyuntil the concentration of ethanol was 50%, and allowed to sitovernight. The supernatant was collected and the precipitates werediscarded. The supernatant was concentrated and mixed with 200 g filler.The mixture was then put in the granulator, added 60 ml of 30 wt. %ethanol, formed into pellets, and dried.

Second, 50 g of vitamins were mixed with 120 g of filler. The mixturewas put into the granulator, added 35 ml of 6 wt. % adhesive, formedinto pellets, and dried.

Third, the above two types of pellets were mixed in prescriptionproportion, tested, formed into tablets in a 33 rotary type tabletpresser, and packed.

Example 7 Preparation of Pellets of the Chinese Traditional MedicineComposition of the Present Invention

First, the following herbs were obtained: 918 g of herba houttuyniae,765 g of caulis lonicerae japonicae, 612 g of radix isatidis, 459 g ofmale fern rhizome, 306 g of radix et rhizoma sophorae tonkinensis, 306 gof radix angelicae dahuricae, 306 g rhizoma paridis, 306 g of herbaartemisiae annuae, and 306 g of rhizoma iridis tectori. The herbahouttuyniae was boiled in water for 40 minutes, filtered, concentratedto a clear cream with a density of 1.05, cooled, added with ethanoluntil the amount of ethanol was 80%, sat for 48 hours, and filteredagain. Ethanol was recovered from the filtrate, concentrated to a clearcream with a density of 1.05. The rest of the 8 herbs were boiled inwater twice for 2 hours each time and filtered. The filtrate wasconcentrated to a clear cream with a density of 1.08, filtered andconcentrated to a density of 1.10. Then, the two clear creams were mixedtogether and concentrated continuously until a thick cream with adensity of 1.26 or above was obtained. The thick cream and powdersucrose were mixed in a proportion of 1:5, formed into pellets, andpacked.

Example 8 Preparation of Capsules of the Chinese Traditional MedicineComposition of the Present Invention

First, the following herbs were obtained: 200 g of herba houttuyniae,500 g of caulis lonicerac japonicae, 384 g of radix isatidis, 800 g ofmale fern rhizome, 200 g of radix et rhizoma sophorae tonkinensis, 200 gof radix angelicae dahuricae, 1000 g rhizoma paridis, 1000 g of herbaartemisiae annuae, and 200 g of rhizoma iridis tectori. All the herbswere added to 3200 ml of injection water, soaked for 1 hour, mixed with7200 ml of injection water, boiled twice for 1 hour each time, andfiltered. The filtrates were combined, recovered under vacuum until nomore ethanol remained, concentrated into a thick cream with a density of1.0 (tested at 60° C.). The cream was put in a vacuum drier, dried at70° C. under vacuum for 36 hours, then crushed and sieved in 60-80 mesh.The medicine powder was then mixed with accessory agents, sieved again,and made into pellets. The pellets were dried, sieved in 20 mesh, mixedwith magnesium stearate, and made into capsules.

Example 9 Preparation of Capsules of the Chinese Traditional MedicineComposition of the Present Invention

First, the following herbs were obtained: 600 g of herba houttuyniae,2000 g of caulis lonicerae japonicae, 1200 g of radix isatidis, and 484g of radix et rhizoma sophorae tonkinensis. All the herbs were added to3200 ml of injection water, soaked for 1 hour, mixed with 3200 ml ofinjection water, boiled twice for 2.5 hours each time, and filtered. Thefiltrates were combined, recovered under vacuum until no more ethanolremained, and concentrated into a thick cream with a density of 1.30.The cream was put in a vacuum drier, dried at 80° C. under vacuum for48-96 hours, then crushed and sieved in 60-80 mesh. The medicine powderwas then mixed with accessory agents, sieved again in 60 mesh, andformed into pellets. The pellets were then dried, sieved in 20 mesh,mixed with magnesium stearate, and made into capsules.

Example 10 Preparation of Milk Products (Health Foods) of the ChineseTraditional Medicine Composition of the Present Invention

First, the following herbs were obtained: 200 g of herba houttuyniae,2000 g of caulis lonicerae japonicae, 584 g of radix isatidis, and 1500g of radix et rhizoma sophorae tonkinensis. All the herbs were added to3200 ml of injection water, soaked for 1 hour, mixed with 3200 ml ofinjection water, boiled twice with 2.5 hours each time, and filtered.The filtrates were combined, recovered under vacuum until no moreethanol remained, concentrated into a thick cream with a density of1.08.

The cow's milk was tested, pasteurized and preprocessed according tostandard (not for skim milk). 0.5-6 mg of the above concentration ofsolution was added to every 100 g of milk. This was then added withcertain flavor. The milk mixtures were filtered, preheated (≦115° C.),pasteurized, homogenized (not for skim milk), cooled, added with yeasts(canned first if it would solidify), and allowed to ferment. Thesolidified, fermented milk was then crushed (not for solidifying type),cooled, canned (not for solidifying type), and tested to obtain thefinal product.

Example 11 Preparation of Drinks (Food) of the Chinese TraditionalMedicine Composition of the Present Invention

First, the following herbs were obtained: 600 g of herba houttuyniae,1000 g of caulis lonicerae japonicae, 1800 g of radix isatidis, 80 g ofmale fern rhizome, and 404 g of radix et rhizoma sophorae tonkinensis,200 g of radix angelicae dahuricae, 100 g of dandelion, and 100 g ofherba andrographics. All herbs were thoroughly washed, crushed, andadded to base solution containing 2%-3% of NaOH (based on the weight ofdrug) and 0.3% of sodium hexametaphosphate (based on the weight ofdrug). The solution was heated at 85° C. for 5 minutes and washed inrunning water until neutral. Second, the medicine was cut in thickflakes, made into a thick liquid in an electrical liquefier, allowed tosit in a stainless steel container, combined with 2% edible phosphate,then heated in an 75° C. water bath for 70 minutes with constantstirring. Third, the thick liquid was neutralized with edible sodiumbicarbonate, while the liquid was cooled in water bath with constantstirring. The fluid was then filtered, pressed and dried. Fourth, thefiltrate was added with 6% of powder active carbon, incubated in an 88°C. water bath for 120 minutes, and filtered. Slightly yellow or brownfiltrate was obtained. The filtrate was then pasteurized, packed andmixed with hydrolyzed medicine, phosphate, and citrate in a proportionof 100:3:4. The medicine was added with water in a proportion of 1:50.Finally the medicine was mixed, packed in bottles, sealed andpasteurized.

Example 12 Preparation of Medicine of Chinese Traditional MedicineComposition of the Present Invention for Animals

First, the following herbs were obtained: 400 g of herba houttuyniae,400 g of caulis lonicerae japonicae, 200 g of radix isatidis, 100 g ofmale fern rhizome, and 1000 g of radix et rhizoma sophorae tonkinensis,1000 g of radix angelicae dahuricae, 1000 g of rhizoma paridis, 1000 gof herba artemisiae annuae, and 1000 g of rhizoma iridis tectori. Allherbs were crushed, filtered through 80 mesh, mixed, and packed.

Example 13 Preparation of Vaccines of the Chinese Traditional MedicineComposition of the Present Invention

First, the following herbs were obtained: 1500 g of herba houttuyniae,400 g of caulis lonicerae japonicae, 584 g of radix isatidis, 800 g ofmale fern rhizome, and 500 g of radix et rhizoma sophorae tonkinensis,and 500 g of radix angelicae dahuricae. All herbs were boiled in waterand filtered. The filtrate was concentrated under vacuum, combined withethanol, filtered, and concentrated again under vacuum to a density of11 g drug/ml. The solution was diluted in water, stored in a fridge,filtered, concentrated a third time under vacuum, and separated inmacroporous resin which was washed subsequently with water, NaOH, and 55wt % ethanol. The medicine was washed using 65 wt % ethanol. The eluatewas collected, diluted in water, combined with L-Arginase, adjusted to apH of 7.0, combined with active carbon, and filtered. The final vaccinewas obtained by mixing the same volume of the filtrate withphysiological saline.

Example 14 Preparation of Capsules of the Chinese Traditional MedicineComposition of the Present Invention

First, the following herbs were obtained: 2000 g of herba houttuyniae,200 g of caulis lonicerae japonicae, 400 g of radix isatidis, 900 g ofradix et rhizoma sophorae tonkinensis, and 784 g of radix angelicaedahuricae All herbs were soaked in 3200 ml of water for 1 hour, boiledin 3200 ml of injection water twice for 2.5 hours each time, andfiltered. The filtrate was then combined, recollected under vacuum untilno more ethanol remained, and concentrated into a thick cream with adensity of 1.30. The cream was dried in a vacuum oven at 80° C. for48-96 hours, crushed, and filtered through 60-80 mesh. The powdermedicine was mixed with accessory agent, filtered through 60 mesh,formed into pellets, dried, combined with magnesium stearate, and madeinto capsules.

Example 15 Preparation of Preventive Injections of the ChineseTraditional Medicine Composition of the Present Invention

First, the following herbs were obtained: 484 g of herba houttuyniae,500 g of caulis lonicerae japonicae, 1800 g of radix isatidis, and 1500g of radix et rhizoma sophorae tonkinensis. All herbs were boiled inwater, filtered, concentrated under vacuum, combined with ethanol, andfiltered again. The filtrate was concentrated under vacuum to a densityof 11 g drug/ml, diluted in water, stored in a fridge, filtered, andconcentrated a third time under vacuum. The concentrate was separated inmacroporous resin which was washed subsequently with water, NaOH, and 55wt % ethanol. The medicine was washed off using 65 wt % ethanol. Theeluate was collected, diluted in water, combined with L-Arginase,adjusted to a pH of 7.0, combined with active carbon, and filtered. Thefinal drug was obtained by mixing the same volume of the filtrate withphysiological saline.

Example 16 Preparation of Capsules of the Chinese Traditional MedicineComposition of the Present Invention

First, the following herbs were obtained: 1600 g of herba houttuyniae,800 g of caulis lonicerae japonicae, 800 g of radix isatidis, 600 g ofradix et rhizoma sophorae tonkinensis, and 484 g of herba andrographis.All herbs were soaked in 3200 ml of injection water for 1 hour, boiledin 3200 ml of injection water twice for 2.5 hours each time, andfiltered. The filtrate was then combined, recovered under vacuum untilno ethanol remained, and concentrated to a thick cream with a density of1.30. The cream was dried in vacuum oven at 80° C. for 48-96 hours,crushed, and filtered through 60-80 mesh. The powder medicine was mixedwith an accessory agent, filtered through 60 mesh, made into pellets,dried, combined with magnesium stearate, and made into capsules.

Example 17 Preparation of Capsules of the Chinese Traditional MedicineComposition of the Present Invention

First, the following herbs were obtained: 1700 g of herba houttuyniae,1000 g of caulis lonicerac japonicae, 400 g of radix isatidis, 500 g ofradix et rhizoma sophorae tonkinensis, and 684 g of Java Brucea Fruit.All herbs were soaked in 3200 ml of injection water for 1 hour, boiledin 3200 ml of injection water twice for 2.5 hours each time, andfiltered. The filtrate was then combined, recovered under vacuum untilno more ethanol remained, and concentrated into a thick cream with adensity of 1.20. The cream was dried in vacuum oven at 80° C. for 48-96hours, crushed, and filtered through 60-80 mesh. The powder medicine wasmixed with accessory agents, filtered through 60 mesh, formed intopellets, dried, combined with magnesium stearate, and made intocapsules.

Example 18 Preparation of Capsules of the Chinese Traditional MedicineComposition of the Present Invention

First, the following herbs were obtained: 1500 g of herba houttuyniae,1000 g of caulis lonicerae japonicae, 100 g of radix isatidis, 800 g ofmale fern rhizome, 500 g of radix et rhizoma sophorae tonkinensis, and384 g of radix angelicae dahuricae. All herbs were soaked in 3200 ml ofinjection water for 1 hour, boiled in 7200 ml of injection water twicefor 1 hour each time, and filtered. The filtrate was then combined,recovered under vacuum until no ethanol remained, and concentrated intoa thick cream to a density of 1.0 (tested at 60° C.). The cream wasdried in a vacuum oven at 70° C. for 36 hours, crushed, and filteredthrough 60-80 mesh. The powder medicine was mixed with accessory agents,filtered through 20 mesh, formed into pellets, dried, combined withmagnesium stearate, and made into capsules.

Example 19 Preparation of Syrup of the Chinese Traditional MedicineComposition of the Present Invention

First, the following herbs were obtained: 200 g of herba houttuyniae,200 g of caulis lonicerac japonicae, 1800 g of radix isatidis, and 1500g of radix et rhizoma sophorae tonkinensis. Naphtha was extracted fromthe herbs by steam. The herbs were then boiled in 6˜8 times of theirvolume of water for 3 times, 1 hour each time. The solution wascombined, filtered, concentrated under vacuum (40° C.), and precipitatedat low temperature for 24 hours. The supernatant was combined withnaphtha, Syrupus Simplex, preservation (sodium benzoate), and distilledwater until 1000 ml. The solution was mixed, packed and pasteurized inflowing steam for 30 minutes.

1. A method of preparing a Chinese traditional medicine composition fortreatment and prevention of Avian Influenza comprising activeingredients and/or medical acceptable vectors, wherein the said activeingredients include the following raw materials: herba houftuyniae 4.7wt. %-70.0 wt. %, caulis lonicerae japonicae 4.7 wt. %-46.7 wt. %, radixisatidis 2.3 wt. %-42.0 wt. %, radix et rhizoma sophorae tonkinensis 1.2wt. %-35.0 wt. % and other herbs for removing fever and detoxification,wherein the method comprises the steps of: (1) all raw materials areweighed; (2) the herba houttuyniae is boiled in water for 10˜40 minutes,and filtered, then the filtrate is concentrated to obtain a clear creamwith a density of 1.05, and cooled; (3) the clear cream from step 2 isadded with ethanol until the amount of ethanol is 50˜80%, allowed to sitfor 24 hours and filtered, then ethanol is recovered from the filtrateand concentrated to obtain a clear cream with a density of 1.25; (4)other raw materials are boiled in water twice for 2 hours each time andfiltered, then the filtrate is concentrated to a density of 1.20,filtered, and concentrated again to obtain a clear cream with a densityof 1.10-1.25; (5) the clear cream from step 3 and step 4 are mixed,concentrated continuously to obtain a thick cream with a density of 1.26or above, and added with accessory agents.
 2. The method of claim 1,wherein said other herbs of said Chinese traditional medicinecomposition are at least one of Male Fern Rhizome, Radix angelicaedahuricae, Herba Andrographic, Dandelion, Natural Indigo,Sargentgloryvine Stem and Chinese Licorice.
 3. The method of claim 1,wherein said herba houttuyniae is present in an amount of 4.7 wt. %-58.4wt. %, and wherein said active ingredients further include thefollowing: male fern rhizome 1.9 wt. %-37.3 wt. % and radix angelicaedahuricae 1.2 wt. %-35.0 wt. %, and wherein said other herbs of saidChinese traditional medicine composition are at least one of rhizomaparidis, herba artemisiae annuae, Java brucea fruit, herba andrographis,dandelion, mint, platycodon grandiflorus, and Chinese licorice.